Phylogenetic conservation of arylsulfatases. cDNA cloning and expression of human arylsulfatase B.
A 2.2-kilobase cDNA clone for human arylsulfatase B (ASB) and several genomic clones were isolated and sequenced. The deduced amino acid sequence of 533 amino acids contains a 41-amino acid N-terminal signal peptide and a mature polypeptide of 492 amino acid residues. Overexpression of ASB in transfected baby hamster kidney (BHK) cells resulted in up to 68-fold higher ASB activity than in untransfected BHK cells. Pulse-chase labeling showed that ASB was synthesized and secreted as a 64-kDa precursor and processed to a 47-kDa mature form in BHK cells. The 47-kDa ASB form was located in dense lysosomes. Transport of ASB to the lysosomes was accomplished in a mannose 6-phosphate receptor-dependent manner. The ASB cDNA clone hybridizes to 4.8-, 2.5-, and 1.8-kilobase species of RNA from human fibroblasts. The same pattern was observed in RNA from fibroblasts of three Maroteaux-Lamy patients who were deficient in ASB activity, as well as in RNA from fibroblasts of three patients with multiple sulfatase deficiency, in which all known sulfatases were markedly diminished. Deduced amino acid sequences of human arylsulfatase A, human ASB, human steroid sulfatase, human glucosamine-6-sulfatase, and an arylsulfatase from sea urchin showed a substantial degree of similarity suggesting that they arose from a common ancestral gene and are members of an arylsulfatase gene family.
Peters
C
C
Biochemie II, Universität Göttingen, Federal Republic of Germany.
Schmidt
B
B
Rommerskirch
W
W
Rupp
K
K
Zühlsdorf
M
M
Vingron
M
M
Meyer
H E
HE
Pohlmann
R
R
von Figura
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001192:Arylsulfatases / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line; D002803:Chondro-4-Sulfatase / Q000235:genetics*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D015870:Gene Expression; D005796:Genes; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes; D010446:Peptide Fragments / Q000032:analysis; D010802:Phylogeny; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D013429:Sulfatases / Q000235:genetics*; D014357:Trypsin
D015345:Oligonucleotide Probes; D010446:Peptide Fragments; D004247:DNA; D013429:Sulfatases; D001192:Arylsulfatases; D002803:Chondro-4-Sulfatase; D014357:Trypsin